Purpose To establish a new method for detecting p16INK4a in cervical tissues with time-resolved fluoroimmunoassay (TRFIA).Methods 126 cases of paraffin imbedding tissues of cervix were selected for immunohistochemistry (IHC) of EnVision two-step and TRFIA.Results There were 20 cases of no intraepithelial lesion or malignancy,24 cases of low-grade squamous intraepithelial lesion (LSIL),53 cases of high-grade squamous intraepithelial lesion (HSIL) and 29 cases of squamous cell carcinoma (SCC).In the groups of no intraepithelial lesion or malignancy,LSIL,HSIL and SCC,p161NK4a positive was seen in 1,19,53 and 28,respectively.TRFIA test results displayed p16INK4a positive in 3,17,50 and 27 cases,respectively.Positive of p16 using by TRFIA in no intraepithelial lesion or malignancy,LSIL,above HSIL was 15.00％,70.83％ and 93.90％,respectively (P ＜ 0.01).Conclusion TRFIA is suitable for detecting of p16INK4a protein and demand low detection equipment,p16INK4a expression detected by TRFIA may helpful for large scale detection in various clinical institution.

Objective To evaluate the correlation between thin-section CT morphologic features and pathological vascular invasion in the clinical stage ⅠA peripheral lung adenocarcinoma.Methods The preoperative thin-section CT images,clinical and pathological characteristics of 102 nodules in 98 patients were retrospectively analyzed and divided into two groups according to pathological vascular invasion.Variables including the diameter of tumor(T) and consolidation part (C) in the mix ground glass nodule (mGGN),C/T ratio,morphology and pathological classification were analyzed by two-independent samples Mann-Whitney U test or t test and x2 test or Fisher exact probability test.Variables with significant difference in the univariate analyses were entered into multivariate analysis to explore predictors for vascular invasion.The correlation between these measurements and vascular invasion was evaluated by a ROC analysis.Results All of 102 nodules,36 had pathological vascular invasion and 66 without.The numbers of two groups with speculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation were 30,30;26,26;35,3;35,39;32,39 and the mediandiameter of tumors were 2.1(1.0-3.0),1.2(0.5-3.0) cm respectively.By univariateanalysis,spiculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation,and diameter of tumor were significantly associated with pathological vascularinvasion (P＜0.01).By multivariate logistic analysis,the statistically significant difference were found in speculation and abnormal vein (OR=33.867,8 161.063,P＜0.05).The ROC analysis showed the area under curve,cut-off value,sensitivity and specificity ofthe diameter of tumor and consolidation part of mGGN and C/T ratio were 0.784,2.05 cm,66.7％,78.8％;0.886,1.08 cm,82.4％,90.0％ and 0.861,0.65,79.4％,85.0％ respectively in differentiating tumors with pathological vascular invasion.Conclusion The spiculation sign and abnormal vein are independent predictors for vascular invasion in stage ⅠA peripheral lung adenocarcinoma.

Objective To analyze the value of predict lymph node status according to the tumor size, solid size, and maximum standardized uptake value(SUVmax) in adenocarcinoma of the lung with a size of ≤3 cm. Methods One hundred and thirty-six patients who had undergone curative resection for lung adenocarcinoma were enrolled in this retrospective study.The largest transverse sectional diameter of the tumor and solid size were measured. Then SUVmax of the main tumor was assessed. Logistic regression analysis was performed by taking the lymph node status as the dependent variable,and the size of solid component of tumor, SUVmax were taken as independent variables. The predicted probability of the Logistic regression model was enrolled in receiver operating characteristic analysis.Results Among the 136 cases of lung adenocarcinoma, the incidence of lymph node metastasis was 24.3%(33/136). Logistic regression analysis showed that the size of the solid component (OR 3.880, 95%CI 1.788 to 8.421, P<0.05) and SUVmax (OR 1.179, 95%CI 1.024 to 1.357, P<0.05) at baseline predicted the metastasis of lymph node independently. And the size of tumor was non-independent predict factor (OR 1.238, 95%CI 0.360-4.260, P>0.05). The ROC curve analysis was performed based on the predicted probability of Logistic regression model, and the area under the curve was 0.838(P<0.01) Conclusions Solid size and SUVmax are important predictors for lymph node metastasis of lung adenocarcinoma with a size of ≤3 cm, and it may be helpful to avoid unnecessary limited resection or lymph node dissection.

Objective To investigate the predictive value of the whole nodule size and solid component size of lung adenocarcinoma manifesting as subsolid nodule(SSN) in three different dimensions for pathologic grade.Methods We evaluated retrospectively preoperative chest HRCT data of 125 patients with 127 SSNs surgically resected and pathologically conformed lung adenocarcinomas.All specimens were divided into two groups: a total of 69 SSNs in group A, including 22 AIS and 47 MIA;a total of 58 SSNs in group B, only including IAC.Computer aided diagnosis software were used to measure the one dimension maximum diameter of solid component with lung window setting(1D-SCLW),two dimension maximum diameter of solid component with lung window setting(2D-SCLW),one dimension maximum diameter of solid component with mediastinal window setting(1D-SCMW),two dimension maximum diameter of solid component with mediastinal window setting(2D-SCMW),one dimension maximum diameter of whole nodule with lung window setting (1D-WNLW), two dimension maximum diameter of whole nodule with lung window setting (2D-WNLW), and volume of solid component with threshold of-300 HU (SCT) of all SSNs.Results 1D-SCLW, 2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW,2D-WNLW and SCT of the group B were significantly larger than those of the group A(P=0.000).ROC analyses indicated that the diagnostic efficiency of SCT for the pathologic grade was the highest among 7 CT features(AUC=0.887, sensitivity:81%,specificity:93%);The cut-off values of 1D-SCLW,2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW, 2D-WNLW and SCT were 17.50 mm,14.75 mm,9.50 mm,7.75 mm,0.50 mm,1.25 mm and 139.00 mm3.Multiple Logistic regression analysis revealed that SCT was the independent predictor of pathologic grade(OR=4.978,95%CI=1.430-17.331,P=0.012).SCT of 139.00 mm3 or greater was a significant indicator of IAC.Conclusion Among the whole nodule size and solid component size of SSN in three different dimensions on preoperative HRCT, SCT is found to be the independent predictor of pathologic grade, which may provide reference for surgery.

Objective To investigate the predictive value of whole nodule size and solid component size of pulmonary subsolid nodules (SSNs)with different window setting on preoperative HRCT for pathologic grade in lung adenocarcinoma.Methods We retrospectively evaluated preoperative chest HRCT and pathological data of 125 patients with 127 surgically resected lung adenocarcinoma manifesting as SSNs.All specimens were divided into two groups:a total of 69 SSNs in group A,including 22 adenocarcinomas in situ (AIS) and 47 minimally invasive adenocarcinoma (MIA);a total of 58 SSNs in group B,including invasive adenocarcinoma (IAC).Observer 1 used computer aided diagnosis software to measure the volume of whole nodule with lung window setting (WNLW),volume of solid component with lung window setting (SCLW),volume of solid component with mediastinal window setting (SCMW) and volume of solid component with threshold of-300 HU(SCT) of all SSNs.Observer 2 randomly selected 50 SSNs and repeated all the measurements.The interobserver agreement regarding quantitative measurements were evaluated by using intraclass correlation coefficient(ICC).The differences of all quantitative features between two groups were evaluated by Mann-Whitney U test.All the quantitative features were evaluated by using univariate logistic regression analysis,significant quantitative features identified by univariate logistic regression analysis were included in the multivariate logistic regression and independent predictors of pathological grade were obtained.Receiver operating characteristic analysis was conducted for the independent predictive factors that exhibited statistically significant differences in the multivariate logistic regression.Results The interobserver agreement regarding quantitative features were excellent (ICC＞ 0.75).The WNLW,SCLW,SCMW and SCT of group B were significantly larger than those of group A (P＜ 0.001).The univariate logistic regression analysis indicated that WNLW,SCLW,SCMW and SCT were significant (P＜0.001),the multivariate logistic regression analysis indicated that SCT was the independent predictive factor (OR=1.013,95％CI:1.006—1.020,P＜0.001).When SCT larger than 139.00 mm3,SSN was significantly associated with IACs (AUC=0.887,sensitivity=81％,specificity=93％).Conclusion SCT of SSNs on preoperative HRCT can be used to distinguish between AIS-MIA and IAC,which may provide information for choice of operation.

Objective To evaluate the performance of bone suppression images on the detection of lung nodules in comparison with the radiologists'reading results.Methods There were 141 standard posteroanterior digital chest radiographs,which included 95 patients with a solitary nodule and 46 controls.In this observational study,4 observers,including 2 radiologists and 2 residents,in-dicated their confidence level regarding the presence of a nodule for each lung,first by use of standard images,then with the addition of bone suppression images.Receiver operating characteristic (ROC)curve analysis was used to evaluate the observers'performance. Results Average nodule size was (1.9±1)cm (range from 0.9 cm to 2.9 cm).The mean value of the area under the ROC curve (AUC)was significantly improved from 0.844 with use of standard images alone to 0.873 with use of bone suppression images (P<0.01).Conclusion The use of bone suppression images can improve radiologists'performance on detection of lung nodules on chest radiographs.

Objective To investigate the clinical effect of transplantation of the free anterolateral thigh flap and iliotibial band for the repair of soft tissue defects at foot and ankle and functional reconstruction.Methods From January,2008 to January,2013,25 pediatric patients with soft tissue defects at foot and ankle were transplantation of the free anterolateral thigh flap and iliotibial band.Of the 25 cases,there were 12 cases at heel,8 cases at dorsum of foot,3 cases at medial malleolus and 2 cases at external malleolus.All the cases were accompanied with different degree of fracture or dislocation,and with soft tissue defect such as achilles tendon,extensor tendon of dorsal,collateral ligament of ankle.The length of soft tissue defect which were reconstructed with iliotibial band were 3 cm to 16 cm.The causes of injury:19 cases were crushed by motorcycle or bicycle,4 cases were crushed machines,2 cases were injuryed in traffic accidents.The areas of free anterolateral thigh flaps were 8 cm × 5 cm to 18 cm × 12 cm.All the patients begined to early rehabilitative exercise under the protection of orthosis after 2 weeks of the operation.Results All cases were followed up from 6 to 24 months,averaged of 14 months.All the flaps survived,and only 2 cases with necrosis of small area in distal,and which were healed by dressing.The healing time were 12 to 24 days,and the average of 15.1 days; The surgery function were assessed according to Thermann's scale,and the results was 14 cases for excellent,9 cases was good,the general was 2 cases,and the excellent and good rate was 92％.Conclusion The transplantation of the free anterolateral thigh flap and iliotibial band for the repair of soft tissue defect at foot and ankle and functional reconstruction is a safe and effective strategy,and it has the advantages such as covering the wound at foot and ankle approvingly,reconstruction of power device once,the flap and iliotibial band were in the same wound,the trauma of doner site is small invasive,early recovery functional exercise,shorten the treatment cycle,and relieve the suffering of children.

Objective To study the effect of TLR4 activation with LPS on BMP9-induced osteogenic differentiation of immortalized mouse embryonic fibroblasts ( iMEFs).Methods The activation of TLR4/NF-κB signaling path-way was detected by ICC.iMEFs were treated with LPS,BAY11-7082,Adnovirus GFP and BMP9.The early osteo-genic differentiation capability of iMEFs was detected by ALP staining and quantitative assay .The later osteogenic differentiation capability was detected by alizarin red S staining .The expression of later osteogenic differentiation marker gene OCN and OPN were detected by PCR and Western blot .The change of p-Smad1/5/8 was detected by Western blot.The expression of Runx2 and Dlx5 were detected by PCR and Western blot .Results LPS can effec-tively stimulate TLR4/NF-κB signaling pathway .TLR4 activation inhibited BMP 9-induced osteogenic differentiation . BMP9-induced osteogenic differentiation related gene and Smad 1/5/8 signaling activation were inhibited by TLR4 activation .The inhibition effect was partly reversed by BAY 11-7082 ( P<0.05 ) .Conclusions TLR4 activation with LPS can inhibit BMP9-induced osteogenic differentiation of iMEFs cells via NF-κB signaling pathway .

Objective To evaluate the effect of CT temporal subtraction(TS) for detection of lung nodules.Methods 80 cases of CT images (current and previous CT images) and corresponding CT TS images were presented, which included 30 cases with nodules(75 nodules) and 50 controls.4 observers, including 2 radiologists and 2 residents, indicated their confidence level regarding the presence of a new emerging or larger or solid component than before,which first used standard CT images, then with the addition of CT TS images.Receiver operating characteristic(ROC) curve analysis was used to evaluate the observers'' performance.Results The mean value of the area under the ROC curve (AUC) were 0.860 and 0.925 for four observers without and with TS images,respectively(P<0.01).Average sensitivity for detection of lung nodules was improved from 77.3%(58/75) to 89.3%(67/75) by using CT TS images.Conclusion The use of CT TS images can significantly improve radiologists'' performance for detection of lung nodules,especially when a small size solid or subsolid nodule presents near the pulmonary hilum.CT TS is more helpful for residents.

Objective: To investigate the awareness of air pollution protection knowledge and its influencing factors among primary school students in Shennongjia Forest District, Hubei Province, so as to provide insights into targeting implementation of health education on air pollution protection knowledge. Methods: Students in Grade 3 to 5 in Shennongjia Shiyan primary school were enrolled by stratified cluster sampling method, and students' demographic features and awareness of air pollution protection knowledge were investigated using the Investigation on the Effects of Air Pollution Health Protection of Pupils (Volume A). Factors affecting the awareness of air pollution protection knowledge among primary school students were identified using a multivariable logistic regression model. Results: A total of 897 questionnaires were allocated, and 877 valid questionnaires were recovered, with an effective rate of 97.77%. The respondents included 446 men (50.86%) and 431 women (49.14%), 301 third grade students (34.32%), 284 fourth grade students (32.38%), and 292 fifth grade students (33.30%), and had a mean age of (10.32±0.93) years. The overall awareness of air pollution protection was 55.76%, and the awareness rates of basic concepts, basic knowledge, and basic behaviors and skills were 42.99%, 53.48% and 57.24%, respectively. Multivariable logistic regression analysis identified age (OR=1.453, 95%CI: 1.053-2.005), living with parents (OR=2.638, 95%CI: 1.571-4.429), mother's educational level (below primary school, OR=0.270, 95%CI: 0.084-0.862; primary school, OR=0.169, 95%CI: 0.069-0.416; junior high school, OR=0.309, 95%CI: 0.138-0.691; high school, OR=0.352, 95%CI: 0.160-0.773) and average annual family income (50 000 to 100 000 Yuan, OR=1.629, 95%CI: 1.162-2.282; 100 000 to 150 000 Yuan, OR=1.802, 95%CI: 1.101-2.948; ≥150 000 Yuan, OR=1.939, 95%CI: 1.065-3.529) as factors affecting the awareness of air pollution protection knowledge among primary school students. Conclusion The awareness of air pollution protection knowledge is 55.76% among primary school students in Shennongjia Forest District, and is affected by age, mother's educational level, average annual family income and living with parents.